不同能量激光照射后中华大蟾蜍早期胚胎发育及其蛋白质谱变化的研究

应用YAG倍频激光器选择不同能量密度激光微束对中华大蟾蜍4细胞期胚胎进行照射,观察胚胎的生长、发育状况,统计出膜率、畸胎率和死亡率。实验结果表明：低能量激光照射明显地提高了胚胎的孵化速度,且使畸变率降低;而高能量激光对胚胎发育有明显抑制作用,而且使畸变率增高。不同能量密度激光微束照射中华大蟾蜍早囊胚,0．5h后制备样品进行双向电泳,凝胶成像后用PDQuest软件进行分析。结果显示激光照射对蛋白质点数及其分布有影响,影响程度与照射激光密度有关。结论：激光照射可以影响胚胎发育和蛋白质的合成。     

He—Ne激光辐照天使鱼胚胎导致畸变

采用He—Ne激光(632.8nm辐照了天使鱼(Pterophyllum eimeki)的发育阶段的胚胎,观察和分析激光辐照对胚胎发育的致变影响和各发育阶段胚胎对激光的受激反应。对受精卵、囊胚期、原肠期和神经胚期的胚胎进行照射,均引起各期胚胎的畸变,这些畸变大多数是初孵仔鱼的头部、尾部、胸腔和背脊的变形。实验结果表明,天使鱼各个发育阶段的胚胎畸变率随着激光辐照剂量的改变而发生变化。     

低强度He-Ne激光照射人外周血淋巴细胞两组试验结果的分析

本文采用染色体畸变 (chromosomalaberration CA)试验和微核 (micronucleus)试验两种方法对低强度He Ne激光辐照育龄妇女外周血淋巴细胞。激光能量密度分别为 14.31J cm2 (辐照 5′)、2 8.6 2J cm2 (辐照 10′) ,5 7.2 4J cm2(辐照 2 0′) ,114.5 2J cm2 (辐照 40′)。照射血样后 ,染色体畸变试验检测其淋巴细胞染色体畸变率 ,激光照射及空白对照组 ,血样染色体畸变率分别为 4.2 9‰、3.96‰、3.81、3.5 9‰和 4.19‰ ,X2 检验无显著意义 (P >0 .0 5 )。阳性对照丝裂霉素MMC处理的血样淋巴细胞CA率平均为 14.41‰ ,明显高于激光照射和空白对照组 ,X2 检验有显著差异(P <0 .0 1)。微核试验检测结果 ,微核染色体分别为 1.0 2‰ ,1.17‰ ,1.18‰ ,1.31‰和 1.19‰对照 ,经统计分析激光照射各组与对照组微核率均在 2‰以下 (P >0 .0 5 ) ,属正常人体微核范围内。结果显示两组试验监测诱变均具有一致性。证明He Ne激光辐照人体细胞对染色体无致畸效应。且表明He Ne激光在治疗范围内应用安全、有效、不会对不体造成危害。     

激光显微照射金鱼受精卵对其胚胎发育的影响

2-细胞期胚胎经激光显微照射(功率为90毫瓦)其中一个卵裂球时能产生明显的损伤光斑,受照射的卵裂球立即停止发育。另一未受照射的卵裂球仍能正常卵裂直至孵化幼鱼,所获得的幼鱼在形态上与正常幼鱼相同。8-细胞或囊胚期胚胎经激光显微照射(功率为372毫瓦)时,则可在受照射的卵裂球上产生明显伤斑,并在照射部位溢出部分细胞内含物,绝大部分胚胎发育成不正常的胚体和各种畸形幼鱼。     

γ－射线照射小鼠后F_1代中的染色体异常和显性致死突变研究

本文报道雌鼠及雌雄性小鼠分别受到０．１５－１．５６Ｇｙ~６０Ｃｏγ－射线照射后,在不同时间进行同笼,取其胚胎观察形态、大小等并制备每个胚胎的染色体标本。分析胚胎细胞染色体数目异常、结构畸变及显性致死效应。结果表明,染色体数目异常发生率与剂量呈直线关系。单纯雌鼠受到１．０７Ｇｙ照射后观察到平衡易位携带者的再现率为０．９９％。雌雄鼠均受到１．０７及１．５６Ｇｙ照射后其Ｆ_１子代中平衡易位携带者的再现率皆为０．３２％。雌雄鼠均受照射组中的显性致死率明显高于雌鼠受照射组,且剂量效应关系为直线相关     

γ—射线照射小鼠后F1代中的染色体异常和显性致死突变研究

１文报道雌鼠及雌雄性小鼠分别受到０．１５－１．５６Ｇｔ＾６０ｃｏγ－射线照射后,在不同时间进行同笼,取其胚胎观察形态、大小等并制备每个胚胎的染体标本。分析胚胎细胞染色体数目异常、结构畸变及显性致死效应。结果表明,染色体数目异常发生率与剂量呈直线关系。单纯雌鼠受到１．０７Ｇｙ照射后观察到平衡易位携带者的再现率为０．９９％。雌雄鼠均受到１．０７及１．５６Ｇｙ照射后观察到平衡易位携带者的再现率为０．９９     

激光对青蛙卵作用的生物效应

用He—Ne激光分别照射青蛙的卵子、精子和早期胚胎,在相同的激光剂量条件下,观察不同激光照射时间对胚胎发育的影响。通过电镜观察卵膜的超微结构,分析激光作用于细胞的刺激作用机制。     

Molecular Cell | 首次揭示SCNT胚胎早期发育过程中Cohesin对初级合子基因组激活相关基因表达的阻碍

体细胞核移植(somatic cell nuclear transfer，SCNT)技术可将体细胞重编程为全能细胞，但研究人员对这一过程中染色体3D构象的重塑却知之甚少。清华大学生命科学学院颉伟研究组与华中农业大学动科动医学院苗义良研究组通过少量细胞全基因组染色质构象捕获技术(sisHi-C)检测了小鼠SCNT胚胎和体外受精(in vitro fertilization, IVF)胚胎卵裂期染色体构象变化(2020年6月23日在线发表，doi:10.1016/j.molcel.2020.06.001)。研究发现胚胎在卵裂期发育过程中拓扑相关结构域(topologically associating domain, TADs)相较其供体细胞或MII卵显著减弱，但SCNT胚胎在1-细胞期TADs明显强于相应时期的体外受精胚胎，2-细胞期之后两种胚胎TADs的变化趋同，2-细胞期之后TADs减弱，至8-细胞期TADs重新增强。研究人员进一步在1-细胞期敲低了TADs关键蛋白Cohesin (黏连蛋白)，发现Cohesin敲低的SCNT胚胎1-细胞期TADs和对照组相比明显减弱，胚胎发育至囊胚的比例显著增加。转录组测序发现Cohesin敲低后的染色体构象变化促进了初级合子基因组激活相关基因的表达，从而促进了SCNT胚胎的体外发育。该研究结果表明染色体构象的异常可能是SCNT胚胎发育能力较IVF胚胎低的重要原因，为提高SCNT胚胎发育能力提供了新思路。     

激光辐照蓖麻蚕的效应探讨

本文采用不同激光波长和不同形式的激光照射蓖麻蚕蚕卵,观察蚕卵的孵化率;蚕血液的酯酶同工酶、血清蛋白、染色体等在激光照射前后的变异情况。     

CO2激光对蚕豆诱变效应试验研究

７０年代以来,全国已有２０多个省市,近百个单位开展激光在农业上应用的研究,并对２０多种作物２００多个品种进行激光育种的研究。试验表明：激光在刺激作物生长,早熟增产以及诱发变异、改良品种等都有一定的效果,并培育出一些新品种应用于生产。然而,激光对作物的作用机制,特别是遗传机理方面的研究报道尚少。本试验目的是探讨ＣＯ２激光对蚕豆根尖细胞染色体畸变的影响及田间农艺性状的变异情况,分析细胞染色体畸变与农艺性状变异的相关性及其遗传传递情况,为作物激光诱变育种提供依据。本试验采用功率密度为３９４ｍＷ／ｃｍ２的ＣＯ２激光照射蚕豆萌动种子,观察其对蚕豆根尖细胞染色体畸变及幼苗生长的影响,探讨ＣＯ２激光照射后蚕豆植株农艺性状的变异及其遗传传递情况。试验结果表明：ＣＯ２激光不同时间照射蚕豆萌动种子,均能使根尖细胞产生染色体畸变,且畸变类型相似;同时,对Ｌ１幼苗生长有明显的抑制作用,Ｌ２植株高度、分枝数及结荚数等农艺性状均产生不同程度的变异。其中分枝数这一性状的变异能传递给Ｌ３,其余性状的变异则属生理损伤所致。     

Lethal and sub-lethal impacts of pulsed laser irradiations on the larvae of the fouling barnacle Balanus amphitrite

Laboratory experiments were conducted to study the impact of laser irradiation on the larvae of the fouling barnacle Balanus amphitrite. Research pertaining to fouling invertebrate larvae-laser interaction is sparse and, hence, data on this aspect were thought significant in order to consider pulsed low power laser irradiations as a possible future antifouling tool. Lethal and sub-lethal impacts of four very low laser fluences, viz. 0.013, 0.025, 0.05 and 0.1 J cm-2 for three different durations, viz. 2, 10 and 30 s were investigated. Three growth stages of barnacle larvae, viz. nauplii stage II, nauplii stage IV and cyprids were exposed to the mentioned laser fluences for different durations. While lethal impact was assessed immediately after and 1 d after irradiation, sub-lethal impacts were studied by monitoring the success rate of the irradiated nauplii in reaching the cyprid stage. In addition, the swimming speed of VIth stage nauplii after irradiation was studied. In the case of cyprids, in addition to the mortality measurement immediately after and 1 d after irradiation, the settlement rate was investigated. In all the above experiments, non-irradiated larvae served as controls. The results showed an increase in mortality with increasing laser fluence and duration of irradiation. Irradiation for 2 s resulted in significant mortality in nauplii, while it was less in the case of cyprids. In IInd stage nauplii, the mortality immediately after irradiation for 2 s varied from 14.8 +/- 2.12 to 97.1 +/- 4.1% for laser fluences of 0.013 and 0.1 J cm-2, respectively. However, in cyprids, the mortality immediately after irradiation for 2 s varied from 12.2 +/- 3 to 13.4 +/- 1.2% for fluences of 0.013 and 0.1 J cm-2, respectively. The mortality in IVth stage nauplii was less than that for IInd stage nauplii but more than that for cyprids. There was a significant increase in mortality with time after irradiation. The formation of cyprids from the irradiated larvae was significantly less than that observed for non-irradiated larvae. Also, the irradiated larvae showed a significantly slower swimming speed compared to the control samples. The settlement rate in cyprids was reduced significantly by the laser irradiation. This was true even for the lowest fluence and shortest period of irradiation tested. Thus, the results of the experiment showed that even a low power pulsed laser irradiation of 0.013 J cm-2 for 2 s can cause significant damage to fouling barnacle larvae.     

Influence low-intensity laser irradiation on the ultrastructural organization of loach embryo cells

The ultrastructural organization of loach embryo cells (Misgurnus fossilis L.) at the stage of the first and the tenth embryo divisions was investigated under the influence of low intensity helium-neon laser irradiation of 5 min exposure. It was determined that the effect of laser irradiation led to ultrastructural changes: the shape of mitochondria and multivesicular bodies changes. The obtained results explain the possible mechanism of influence of low-intensity laser irradiation at the cellular level.     

Lethal and sub‐lethal impacts of pulsed laser irradiations on the larvae of the fouling barnacle Balanus amphitrite

Laboratory experiments were conducted to study the impact of laser irradiation on the larvae of the fouling barnacle Balanus amphitrite. Research pertaining to fouling invertebrate larvae‐laser interaction is sparse and, hence, data on this aspect were thought significant in order to consider pulsed low power laser irradiations as a possible future antifouling tool. Lethal and sub‐lethal impacts of four very low laser fluences, viz. 0.013, 0.025, 0.05 and 0.1 J cm^‐2 for three different durations, viz. 2, 10 and 30 s were investigated. Three growth stages of barnacle larvae, viz. nauplii stage II, nauplii stage IV and cyprids were exposed to the mentioned laser fluences for different durations. While lethal impact was assessed immediately after and 1 d after irradiation, sub‐lethal impacts were studied by monitoring the success rate of the irradiated nauplii in reaching the cyprid stage. In addition, the swimming speed of VI^th stage nauplii after irradiation was studied. In the case of cyprids, in addition to the mortality measurement immediately after and 1 d after irradiation, the settlement rate was investigated. In all the above experiments, non‐irradiated larvae served as controls. The results showed an increase in mortality with increasing laser fluence and duration of irradiation. Irradiation for 2 s resulted in significant mortality in nauplii, while it was less in the case of cyprids. In II^nd stage nauplii, the mortality immediately after irradiation for 2 s varied from 14.8±2.12 to 97.1±4.1% for laser fluences of 0.013 and 0.1 J cm^‐2, respectively. However, in cyprids, the mortality immediately after irradiation for 2 s varied from 12.2±3 to 13.4±1.2% for fluences of 0.013 and 0.1 J cm^‐2, respectively. The mortality in IV^th stage nauplii was less than that for II^nd stage nauplii but more than that for cyprids. There was a significant increase in mortality with time after irradiation. The formation of cyprids from the irradiated larvae was significantly less than that observed for non‐irradiated larvae. Also, the irradiated larvae showed a significantly slower swimming speed compared to the control samples. The settlement rate in cyprids was reduced significantly by the laser irradiation. This was true even for the lowest fluence and shortest period of irradiation tested. Thus, the results of the experiment showed that even a low power pulsed laser irradiation of 0.013 J cm^‐2 for 2 s can cause significant damage to fouling barnacle larvae.     

激光照射对中华大蟾蜍原肠胚蛋白质表达的影响

用YAG倍频激光器选择不同能量密度对中华大蟾蜍原肠胚进行照射,对各组样品进行双向电泳,凝胶成像后用PDQuest软件进行分析,结果表明:激光照射对蛋白质点数及其分布影响显著,影响程度与照射激光密度有关。各激光照射组与对照组相比,有些蛋白质点变化明显,有些蛋白质点在对照组未发现,而在各激光照射组皆新出现。结论:激光照射可对原肠胚一些蛋白质的表达产生明显的影响。     

Mitotic reversion in prophase of PTK1 cells induced by argon laser microirradiation

In this paper, we report the effects of laser microirradiation of prophase nucleoli and mitotic chromosomes in cells of female rat kangaroo kidney epithelial cell line PTK1. When the laser power delivered to sample surface was 90-190 mW, irradiation of one of the two nucleoli in the prophase cell did not inhibit the mitotic progress, but resulted in the loss of the irradiated nucleolus in daughter cells. When the laser power was increased to 360-420 mW, either irradiation of the nucleolus or chromosome in midprophase caused a blockage of mitosis at terminal midprophase. The irradiated cells returned morphologically to early prophase. No mitotic reversion occurred in the case of irradiation of chromosomes at late prophase, prometaphase, metaphase, and anaphase. Irradiation of the cytoplasm in prophase cells caused a 50-70 min mitotic delay at prophase. However, the irradiated cells underwent successive mitotic divisions. The mechanism of laser-induced mitotic prophase reversion is discussed.     

Effect of the time before fixation on the yield of chromosome aberrations in a culture of human lymphocytes exposed to gamma rays at different stages of the mitotic cycle

A comparative study was made of the yield of chromosome aberrations in human lymphocyte culture exposed to 60Co-gamma-rays (2 Gy) at different mitotic cycle stages the cells being fixed after 52 and 60 hr. It was shown that with the latter fixation time (60 hr) the frequency of chromosome aberrations after irradiation in G1 stage was substantially lower than that with the former one (52 hr) and, vice versa, it was higher after irradiation in S and G2 stages. The authors discuss the probable causes of the distinctions observed.     

Mitotic reversion in prophase of PTK1 cells induced by argon laser microirradiation

In this paper, we report the effects of laser microirradiation of prophase nucleoli and mitotic chromosomes in cells of female rat kangaroo kidney epithelial cell line PTK₁. When the laser power delivered to sample surface was 90–190 mW, irradiation of one of the two nucleoli in the prophase cell did not inhibit the mitotic progress, but resulted in the loss of the irradiated nucleolus in daughter cells. When the laser power was increased to 360–420 mW, either irradiation of the nucleolus or chromosome in midprophase caused a blockage of mitosis at terminal midprophase. The irradiated cells returned morphologically to early prophase. No mitotic reversion occurred in the case of irradiation of chromosomes at late prophase, prometaphase, metaphase, and anaphase. Irradiation of the cytoplasm in prophase cells caused a 50–70 min mitotic delay at prophase. However, the irradiated cells underwent successive mitotic divisions. The mechanism of laser-induced mitotic prophase reversion is discussed.     

The Irradiation of Dividing Cells: Changes in Sensitivity to X-Rays during Mitosis

Roots of Vicia faba were given combined X-ray and colchicinetreatments. Breaks were not visible immediately after irradiation,even at metaphase. Breaks and reunions were, therefore, scoredlater in 2x nuclei, and in the 4x nuclei derived from 2x nucleiafter colchicine treatment. It was shown that metaphase is themost sensitive stage of mitosis Breaks per chromosome are less frequent in 4x nuclei than in2x nuclei after irradiation of resting stage. This apparentphysiological difference between the two types of nuclei mayaffect either the frequency of breakage or of restitution. Afterthe irradiation of prophase, metaphase, or telophase cells,however, breaks per chromosome are more frequent in the 4x nucleithan they are in 2x nuclei in the same sample owing to the factthat the 2x, unlike the 4x nuclei, are not accurately timed.They must include cells irradiated during the less sensitiveresting stage. Chromosome reunion occurred, probably at telophase. Chromatidreunion and sister reunion did not occur frequently. Chromatidbreaks, like chromatid reunions, were infrequent except afterirradiation of resting stage cells. The evidence indicates thatthe chromosome, at metaphase just as at resting stage, reactsto X-rays as a single functional unit.     

The action of helium-neon laser radiation on the growth of Mycobacterium tuberculosis

The growth properties of M. tuberculosis subjected to the action of helium-neon laser radiation was studied. Laser radiation was shown to change the quantitative and qualitative composition of mycobacterial population. Disturbances in the viability of mycobacteria appear as a consequence of changes in the morphological structure of mycobacterial cells. The maximum effect of helium-neon laser radiation was achieved after the irradiation of M. tuberculosis culture on days 2-3 after inoculation. These results made it possible to suggest that the effect of helium-neon laser radiation was most pronounced in cells at the stage of mitosis (the logarithmic stage of growth) with the highest degree of metabolism.